The present invention relates to a highly effective method, apparatus and kit for growing microorganisms or cell cultures from organisms such as plants or animals, especially derived from culture blocks containing multiple wells, such as a 96-well or 384-well plate or block.
Experiments involving biotechnology include the use of biological systems such as genetically engineered microorganisms or cell cultures. In order to prepare such microorganisms or cell cultures, new genetic material usually in the form of DNA (deoxyribonucleic acid) fragments, is inserted into the microorganisms or cell culture. If successfully inserted, the genetic makeup of the treated microorganisms or cell culture will be changed so that it can express a new trait (e.g. making a new protein). In a traditional method, a sample of a treated microorganisms or cell culture is grown by spreading an aliquot of liquid culture containing the microorganisms or cell culture onto the surface of an individual petri plate containing agar-based medium. Turn-tables and hand-held spreaders to facilitate this process are commercially available (VWR). In this traditional method, however, only one sample at a time can be handled, requiring considerable time, labor and expense to grow the microorganisms or cell cultures on individual plates. Thus, a new approach was sought to develop a high-throughput system and method that would enable one to grow multiple samples of microorganisms or cell cultures, especially from plates or blocks containing multiple wells of microorganisms or cell cultures.
In one embodiment, the present invention is directed towards a method for plating microorganisms or cell cultures, comprising:
a) using an apparatus comprising
i) a tray having a bottom and sidewalls extending from said bottom;
ii) a plurality of substantially uninterrupted parallel dividers, each divider having a length and a height;
iii) means for maintaining said dividers substantially parallel within said tray, wherein said dividers are removable from said tray;
iv) agar that fills said tray to a point less than the height of said dividers in said tray, wherein said agar has a surface for plating microorganisms or cell cultures that is defined as a lane between two adjacent dividers or a lane between a divider and an adjacent sidewall; and
v) beads that can fit between said substantially parallel dividers;
b) contacting said beads with a source of microorganisms or cell culture; and
c) rolling said beads with said microorganisms or cell culture along said agar surface to plate said microorganisms or cell culture.
In another embodiment, the present invention is directed towards an apparatus comprising:
a) a tray having a bottom and sidewalls extending from said bottom;
b) a plurality of substantially uninterrupted parallel dividers, each divider having a length and a height; and
c) means for maintaining said dividers substantially parallel within said tray, wherein said dividers are removable from said tray.
In another embodiment, the present invention is directed towards an apparatus comprising:
a) a tray having a bottom and sidewalls extending from said bottom;
b) a plurality of substantially uninterrupted parallel dividers, each divider having a length and a height;
c) means for maintaining said dividers substantially parallel within said tray, wherein said dividers are removable from said tray; and
d) a cover for covering said tray and said dividers.
In another embodiment, the present invention is directed towards an apparatus comprising:
a) a tray having a bottom and sidewalls extending from said bottom;
b) a plurality of substantially uninterrupted parallel dividers, each divider having a length and a height;
c) means for maintaining said dividers substantially parallel within said tray, wherein said dividers are removable from said tray;
d) a cover for covering said tray and said dividers; and
e) beads that can fit between said substantially parallel dividers.
In another embodiment, the present invention is directed towards an apparatus comprising:
a) a tray having a bottom and side walls extending from said bottom;
b) a plurality of substantially uninterrupted parallel dividers, each divider having a length and a height;
c) means for maintaining said dividers substantially parallel within said tray, wherein said dividers are removable from said tray;
d) a cover for covering said tray and said dividers; and
h) agar that fills said tray to a point less than the height of said dividers in said tray, wherein said agar has a surface for plating microorganisms or cell culture that is defined as a lane between two adjacent dividers or a lane between a divider and an adjacent sidewall.
In another embodiment, the present invention is directed towards an apparatus comprising:
a) a tray having a bottom and sidewalls extending from said bottom;
b) a plurality of substantially uninterrupted parallel dividers, each divider having a length and a height;
c) means for maintaining said dividers substantially parallel within said tray, wherein said dividers are removable from said tray;
d) a cover for covering said tray and said dividers; and
h) agar that fills said tray to a point less than the height of said dividers in said tray, wherein said agar has a surface for plating microorganisms or cell culture that is defined as a lane between two adjacent dividers or a lane between a divider and an adjacent sidewall.
In another embodiment, the present invention is directed towards a kit comprising in a single package:
a) a tray having a bottom and sidewalls extending from said bottom;
b) a plurality of substantially uninterrupted parallel dividers, each divider having a length and a height; and
c) means for maintaining said dividers substantially parallel within said tray, wherein said dividers are removable from said tray.
The method, apparatus or kit of the present invention can utilize or have a plurality of dividers, preferably at least eleven or more dividers. The tray, the dividers, the means for maintaining the dividers parallel (such as supporting ribs), can be plastic. The tray and the dividers can be selected from a plastic, which is polycarbonate, polystyrene, polypropylene or combinations thereof.
The method, apparatus or kit of the present invention can have a means for maintaining the dividers substantially parallel. The means for maintaining the dividers substantially parallel can be at least one supporting rib. The maintaining means can also be at least two, three, four, five or more supporting ribs.
The method, apparatus or kit of the present invention can include a tray, which has an interior length and an interior width and the divider length can be less than the length of the interior of the tray. The divider length can preferably be from about 75% to about 95% of the interior length of the tray. The method, apparatus or kit of the present invention can also include a cover and the cover can be removable from the tray.
The method, apparatus or kit of the present invention can also include beads. Preferably, the beads can be spherical (ball or ball-shaped) or even cylindrical. Also preferred is that the bead are made of glass. In addition, the method, apparatus or kit can also have a starting gate. The dividers used in the present invention can have at least one notch along the edge of the divider length and the starting gate can rest upon the notch in each of the dividers.
The method, apparatus or kit can further have an exit-gate whose removal defines an exit-alley within the tray. The exit-alley can thereby communicate with at least one sidewall of the tray, which can have at least one aperture therein. The method, apparatus or kit can also comprise a tray and dividers that are substantially sterile.
The present invention has the advantage of providing a method, apparatus or kit for plating microorganisms or cell culture that requires significantly less time and fewer personnel to perform compared with other known methods.
Another advantage of the present invention is that it provides a method, apparatus or kit for plating microorganisms or cell culture in a high-throughput manner that can utilize different types of microorganisms or cell cultures on the same plate.